[关键词]
[摘要]
目的 建立白芍药材HPLC指纹图谱及多成分含量测定方法,为白芍药材质量控制及临床应用提供参考。方法KromasiL C18色谱柱(250 mm×4.6 mm,5 μm)为固定相,以乙腈-0.1 %磷酸水溶液为流动相进行梯度洗脱,体积流量1.0 mL/min,检测波长0~17 min和22~45 min,244 nm;17~22 min,280 nm,柱温30 ℃;对16批白芍药材进行指纹图谱构建,基于指纹图谱相似度分析结合聚类分析与主成分分析,同时测定没食子酸、氧化芍药苷、芍药内酯苷、芍药苷、苯甲酰芍药苷的含量。结果 16批白芍药材的相似度均在0.86以上,共标定了9个共有峰,指认出5个色谱峰,分别为没食子酸、氧化芍药苷、芍药内酯苷、芍药苷、苯甲酰芍药苷。聚类分析将16批白芍药材分为2类;经主成分分析,主成分1~3是影响药材样品质量评价的主要因子。16批白芍药材中没食子酸、氧化芍药苷、芍药内酯苷、芍药苷、苯甲酰芍药苷质量分数范围分别为0.09%~0.27%、0.21%~0.35%、0.22%~2.50%、2.74%~5.63%、0.06%~0.28%。经方法学考察,各成分在线性关系良好(r2≥0.999 6),平均回收率为92.07%~100.70%。结论 建立的HPLC指纹图谱及多成分含量测定方法稳定、可靠,可为白芍药材的质量评价提供依据。
[Key word]
[Abstract]
Objective To establish HPLC fingerprints and multi-components determination method of Baishao (Paeoniae Radix Alba), and provide reference for the quality control and clinical application of Paeoniae Radix Alba. Methods Kromasil C18(250 mm×4.6 mm, 5 μm) column was used as stationary phase and acetonitrile-0.1% phosphoric acid aqueous solution was used as mobile phase for gradient elution, with the flow rate of 1.0 mL/min, the detection wavelength was 244 nm at 0—17 min and 22—45 min; 280 nm at 17—22 min and column temperature was 30 ℃. The fingerprints of 16 batches of Paeoniae Radix Alba were constructed. Based on the similarity analysis of fingerprints, cluster analysis and principal component analysis, the content of gallic acid, oxypaeoniflorin, albiflorin, paeoniflorin and benzoylpaeoniflorin were determined. Results The similarity of 16 batches of Paeoniae Radix Alba was more than 0.86, and nine common peaks were identified. The 16 batches of Paeoniae Radix Alba were divided into two categories by cluster analysis. According to the principal component analysis, the principal components 1—3 were the main factors affecting the quality evaluation of the samples. The contents of gallic acid, oxypaeoniflorin, albiflorin, paeoniflorin and benzoylpaeoniflorin in 16 batches of Paeoniae Radix Alba were 0.09%—0.27%, 0.21%—0.35%, 0.22%—2.50%, 2.74%—5.63%, 0.06%—0.28%, respectively. The results showed that the linear relationship between the components was good (r2≥0.999 6), the average recovery was 92.07%—100.7%. ConclusionThe established HPLC fingerprints and multi-components determination method are stable and reliable, which can provide theoretical guidance for the establishment of quality standards of Paeoniae Radix Alba.
[中图分类号]
R286.2
[基金项目]
国家中医药管理局中药标准化项目(ZYBZH-C-JS-30);南京市医学科技发展项目(YKK16173);江苏省科技成果转化专项资金项目(BA2016104)